Swanchara K W, Henricks D M, Birrenkott G P, Bodine A B, Richardson M E
Animal, Diary, and Veterinary Sciences Department, Clemson University, South Carolina 29634-0361, USA.
Biol Reprod. 1995 Oct;53(4):911-22. doi: 10.1095/biolreprod53.4.911.
The production, secretion, and localization of epidermal growth factor (EGF) and the distribution of the EGF receptor (EGF-R) were examined in the isthmus (I) and ampulla (A) of the oviducts from cyclic (C) and early-pregnant (P) gilts. Sexually mature gilts (n = 20) were divided equally into two groups: C and P. P gilts were bred twice (at 0 and 24 h), and all gilts were killed 48 h after onset of estrus. After removal of reproductive tracts, oviducts were isolated, flushed, opened longitudinally, divided by anatomical region, cut into 1-3-mm3 pieces, and placed in Dulbecco's modified Eagle's Essential medium (DMEM: F-12 + ITS [insulin, 5 micrograms/ml; transferrin, 5 micrograms/ml; and selenious acid, 5 ng/ml] + antibiotic). Half the tissue and medium were immediately homogenized and centrifuged, and the supernatant was removed. The remaining tissue was cultured in the medium for 24 h at 37 degrees C and 5% CO2, then prepared similarly for analysis. EGF was measured in the supernatant by a heterologous RIA. Concentration of EGF was expressed as nanogram/milliliter of EGF per milligram of protein in wet tissue. EGF concentrations were present in both regions of the oviducts of C and P gilts. It was greater in I than in A tissues for both C (I = 16.21 ng/ml vs. A = 13.91 ng/ml; p < 0.05) and P gilts (I = 14.27 ng/ml vs. A = 12.53 ng/ml; p < 0.10). Higher concentrations of EGF were found in I tissue of C gilts than in P gilts (C = 16.21 ng/ml vs. P = 14.27 ng/ml; p < 0.05). The media assayed from cultured explants of I and A sections from C and P gilts gave results that were highly correlated with those of immediately prepared tissue sections. Localization of EGF in frozen oviductal tissue sections was demonstrated by immunohistochemistry. The primary site of EGF immunostaining occurred in the epithelial cells (with highest intensity at the apical border) of both C and P gilts. A and I tissue sections from C gilts showed localization of EGF immunostaining mainly in epithelial cells and lamina propria cells, while those from P gilts stained less intensely. The presence of EGF-R was shown by incubating tissue imprints and frozen sections with EGF-erythrosin isothiocyanate, which revealed that EGF-R were distributed mainly on the membranes of epithelial cells. The study indicates that EGF and EGF-R are present in oviductal epithelial cells in both C and P gilts, with the highest concentration of EGF in C gilts.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了发情周期（C）和妊娠早期（P）后备母猪输卵管峡部（I）和壶腹部（A）中表皮生长因子（EGF）的产生、分泌及定位，以及EGF受体（EGF-R）的分布。性成熟后备母猪（n = 20）平均分为两组：C组和P组。P组后备母猪配种两次（0小时和24小时各一次），所有母猪在发情开始后48小时处死。取出生殖道后，分离输卵管，冲洗，纵向切开，按解剖区域分开，切成1 - 3立方毫米的小块，置于杜氏改良 Eagle 基本培养基（DMEM：F-12 + ITS [胰岛素，5微克/毫升；转铁蛋白，5微克/毫升；亚硒酸，5纳克/毫升] + 抗生素）中。一半组织和培养基立即匀浆并离心，去除上清液。剩余组织在培养基中于37℃和5%二氧化碳条件下培养24小时，然后同样制备用于分析。通过异源放射免疫分析（RIA）测定上清液中的EGF。EGF浓度以每毫克湿组织中EGF的纳克/毫升表示。C组和P组后备母猪输卵管的两个区域均存在EGF浓度。C组（I = 16.21纳克/毫升 vs. A = 13.91纳克/毫升；p < 0.05）和P组（I = 14.27纳克/毫升 vs. A = 12.53纳克/毫升；p < 0.10）中，I区的EGF浓度均高于A区。C组后备母猪I区组织中的EGF浓度高于P组（C = 16.21纳克/毫升 vs. P = 14.27纳克/毫升；p < 0.05）。对C组和P组后备母猪I区和A区培养外植体的培养基检测结果与立即制备的组织切片高度相关。通过免疫组织化学证实了冷冻输卵管组织切片中EGF的定位。C组和P组的EGF免疫染色主要位点均位于上皮细胞（顶端边界处强度最高）。C组的A区和I区组织切片显示EGF免疫染色主要定位于上皮细胞和固有层细胞，而P组的染色强度较低。用异硫氰酸荧光素标记的EGF孵育组织印记和冷冻切片显示存在EGF-R，表明EGF-R主要分布在上皮细胞膜上。该研究表明，C组和P组后备母猪输卵管上皮细胞中均存在EGF和EGF-R，C组中EGF浓度最高。（摘要截断于250字）
